Previous studies from this laboratory have shown that low-amplitude electrical activity (LEA) may occur during standard hyperkalemic hypothermic cardioplegic arrest and be undetected by routine monitoring techniques. The present study was designed to elucidate the electrophysiological nature of LEA. Ten dogs were monitored continuously during standard cardioplegic arrest using a 32-channel data acquisition system. In 7 animals (Part I), electrophysiological mapping of the lower right atrial septum during arrest was performed. The initial site of activation of LEA was consistently recorded from the region of the lower atrial septum prior to atrial or ventricular electrical activation. The site of origin of LEA was thus localized to the anatomical region of the atrial septum containing the atrioventricular nodal conduction tissue. In Part II, the electrophysiological mechanism of LEA was investigated in the remaining 3 animals utilizing an intrinsic property of specialized conduction tissue. Inclusion of a calcium channel–blocking agent in standard cardioplegic solution completely prevented the development of LEA in all 3 animals, which is in contrast to findings in previous studies using standard cardioplegic solution alone. These data suggest that LEA may be related to calcium-mediated activation of specialized conduction tissue. This mechanism of activation may explain why LEA cannot be detected by the intraoperative monitoring techniques routinely employed.